This invention relates to crosslinked collagen and glycosaminoglycan materials, and to procedures for preparing such materials. It has been discovered that if collagen fibrils in an aqueous acidic solution are contacted with a crosslinking agent before being contacted with glycosaminoglycan, the materials produced have extremely low levels of thrombogenicity. Such materials are well suited for in-dwelling catheters, blood vessel grafts, and other devices that are in continuous contact with blood for long periods of time.
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2. In the method of preparing crosslinked collagen-glycosaminoglycan composite material, the improvement of contacting collagen, which has been soaked in an aqueous acidic solution, with a crosslinking agent prior to contacting said collagen and said crosslinking agent with glycosaminoglycan.
1. A method for preparing a crosslinked collagen-glycosaminoglycan composite material, comprising:
a. soaking collagen in an aqueous acidic solution having a pH of below about six; b. contacting said solution with a crosslinking agent; and c. contacting a source of glycosaminoglycan with said solution.
5. A method of
6. A crosslinked collagen-glycosaminoglycan composite material made by the methods of
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The invention described herein was supported in whole or in part by a grant from the National Institutes of Health.
This invention is in the fields of polymeric materials and biocompatible materials.
Composite materials made of collagen and glycosaminoglycan (GAG) have been shown to be highly useful for certain biochemical applications. For example, U.S. Pat. No. 4,060,081 (Yannas et al, 1977), the teachings of which are hereby incorporated by reference, discloses a multilayer membrane suitable for use as synthetic skin. The bottom layer, which is placed in contact with a woundbed, is a highly porous lattice comprising collagen that is crosslinked with GAG. This lattice provides a biophysical supporting structure in which cells can migrate and proliferate to heal the wound.
The typical procedures that have been used in the past to prepare collagen/GAG composite materials are described in U.S. application Ser. No. 30,183, filed on Apr. 16, 1979, now U.S. Pat. No. 4,280,954 and U.S. application Ser. No. 169,897, filed on July 17, 1980, now abandoned; the teachings of both of those applications are hereby incorporated by reference. Briefly, a preferred embodiment of those procedures comprises the following steps, in sequence:
1. Mechanically cutting and grinding a source of collagen into particulate form.
2. Soaking the particulate collagen in dilute acetic acid.
3. Homogenizing the solution in a blender.
4. Adding a source of glycosaminoglycan which has been ground into particulate form. Typically, enough GAG is added to the solution to comprise about 6% to about 12% by dry weight of the composite material. The collagen/GAG mixture normally precipitates out of the acidic solution and forms a fibrous dispersion.
5. The precipitate is homogenized in a blender.
6. The solution is contacted with a crosslinking agent such as glutaraldehyde.
7. The solution is quickly frozen in a shallow pan.
8. The frozen dispersion is subjected to a high degree of vacuum, causing the acidic fluid to evaporate while the spatial configuration of the partially crosslinked fibrils is maintained.
The composite material thus formed may be treated by additional procedures to remove all traces of glutaraldehyde and to increase the crosslinking density and strength of the composite material.
The resulting collagen/GAG composite material exhibits some degree of incompatibility with the blood. For example, a small amount of platelet aggregation may occur over a period of several months if blood is in continuous contact with this material. This degree of blood incompatibility might be tolerable if the collagen/GAG composite material is to be biodegraded over a relatively short period of time, as is the case when it is used as artificial skin. However, that level of blood incompatibility can lead to certain problems in the use of collagen/GAG composite materials for prosthetic devices which are not rapidly biodegraded and which are in continuous contact with blood. For example, collagen/GAG material, in a less porous form than the lattice used for synthetic skin, may be useful for in-dwelling catheters, blood vessel replacement material, and other prosthetic devices. For such uses, it is advantageous to utilize a composite material that does not cause platelet aggregation to any detectable extent.
This invention relates to a method of creating collagen and glycosaminoglycan (GAG) composite materials that have extremely high levels of blood compatibility. The Applicants have discovered that if an aqueous dispersion of collagen which has been swollen in acid is contacted with a crosslinking agent such as glutaraldehyde prior to adding GAG to the solution, the resulting composite material will cause extremely low or undetectable levels of blood platelet aggregation. The invention described herein also relates to composite materials formed by this method. These materials are highly useful for prosthetic devices that will remain in contact with blood for very long periods of time, including indwelling catheters, blood vessel grafts, and other prosthetic devices that are not rapidly biodegraded.
In one preferred embodiment of this invention, a source of collagen is ground into particulate form and contacted with dilute acetic acid. The acidic solution is stirred to ensure thorough contact of all collagen with acid. The collagen is subsequently allowed to swell for 72 hours to remove all native banding.
After swelling, a crosslinking agent comprising glutaraldehyde is added in the form of an aqueous solution. The final glutaraldehyde concentration is approximately equal to 0.5% weight/volume. The solution normally is stirred while the glutaraldehyde is being added.
Following the addition of glutaraldehyde, a solution of chondroitin 6-sulfate (which is a glycosaminoglycan) is added to the acidic collagen solution. The chondroitin 6-sulfate is first dissolved in dilute acetic acid, and added dropwise to the collagen solution while it is being stirred.
The resulting dispersion may be treated in any of several ways. In order to create a prosthetic device, it may be frozen and lyophilized. In order to test its blood compatibility properties, it may be filtered, air-dried, ground into particulate form, and dispersed in physiological saline solution.
PAC Example 1: Preparation of Collagen/GAG MaterialCollagen from bovine hide, prepared by the methods described by M. Komanowsky et al, J. Amer. Leather Chemists Assn. 69: No. 9, p. 410-422 (1974), was donated by H. I. Sinnamon of the U.S. Dept. of Agriculture. 0.55 g. of freeze-dried collagen was ground in a Wiley mill (A. H. Thomas Co., Phila. PA) to a 60 mesh particle size. It was added to 200 ml of 0.05 M aqueous acetic acid. This solution was stirred for 60 minutes in an ice-jacketed blender (Eberbach Corp., Ann Arbor, MI) on a 2-speed power unit (Waring Co., Hartford, CT) set on high speed with the line voltage reduced to 50% of 120 volts. Following this blending step, the solution was placed in a tightly closed glass jar and the collagen was allowed to swell for 72 hours at 4°C
A solution of 25% glutaraldehyde ("Baker-analyzed" reagent grade, J. T. Baker Co., Phila. PA) in distilled water was prepared. A sufficient quantity of this solution was added to the acidic collagen solution to comprise 0.5% glutaraldehyde volume per volume. The glutaraldehyde solution was added while the dispersion was blended for one hour in an overhead blender (Hamilton Beach Div. of Scovill, Washington, N.C.) set on the low speed with the line voltage reduced to 60% of 120 volts.
0.0578 g chondroitin 6-sulfate was dissolved in 10 ml of 0.05 M acetic acid. This solution was added to 175 ml of the glutaraldehyde-treated collagen dispersion. The addition was performed over a period of 5 minutes while the dispersion was being blended in the overhead blender.
Shortly thereafter, the dispersion was filtered in a Buechner funnel. This filtering step was completed in about 20 minutes. The resulting wet membrane was then air-dried and milled to a 60 mesh particle size. It was dispersed in physiological saline solution (0.15 M NaCl, pH7) prior to being contacted with blood platelets.
Two other dispersions of collagen/GAG were prepared as described above, with the following changes. In one specimen, the glutaraldehyde crosslinking step was omitted. In the other specimen, chondroitin 6-sulfate was added to the collagen dispersion prior to the addition of glutaraldehyde.
Blood plasma that was rich in platelet content was obtained from healthy, drug-free human donors (Beth Israel Hospital, Boston, MA) by using the procedures described in F. H. Silver et al, "Glycosaminoglycan Inhibition of Collagen-Induced Platelet Aggregation," Thrombosis Research 13: 267-277 (1978). Platelet clotting was measured with an aggregometer (Chrono-Log Corporation Model 300, Broomall, PA).
When platelet-rich plasma was contacted with the collagen/GAG dispersion that had not been contacted with glutaraldehyde, the collagen/GAG material caused rapid formation of a platelet clot. When the platelet-rich plasma was contacted with the collagen/GAG that had been contacted with glutaraldehyde subsequent to the addition of chondroitin 6-sulfate, a small amount of platelet aggregation occurred if the dispersion had been stored for a period of several months in physiological saline prior to being contacted with platelet-rich plasma. By contrast, when the glutaraldehyde crosslinking step preceded the addition of chondroitin 6-sulfate, as described in Example 1, no platelet aggregation could be detected, regardless of whether the dispersion was stored in physiological saline for up to one month prior to being contacted with platelet-rich plasma.
This invention has industrial applicability in the preparation of collagen prosthetic devices that have very high levels of blood compatibility.
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention, and are covered by the following claims.
Yannas, Ioannis V., Forbes, Martin J.
Patent | Priority | Assignee | Title |
10016534, | Nov 17 2008 | PETVIVO HOLDINGS, INC | Protein biomaterial and biocoacervate vessel graft systems and methods of making and using thereof |
10022214, | Dec 20 2011 | LifeCell Corporation | Sheet tissue products |
10077420, | Dec 02 2014 | OCUGEN, INC | Cell and tissue culture container |
10327884, | Jan 24 2012 | LifeCell Corporation | Elongated tissue matrices |
10722339, | Dec 20 2011 | LifeCell Corporation | Flowable tissue products |
10842610, | Jul 03 2007 | OCUGEN, INC | Method for use of a double-structured tissue implant for treatment of tissue defects |
10953044, | Apr 24 2012 | LifeCell Corporation | Functionalized tissue matrices |
11555172, | Dec 02 2014 | OCUGEN, INC | Cell and tissue culture container |
11890371, | Dec 26 2007 | PETVIVO HOLDINGS, INC | Biocompatible protein-based particles and methods thereof |
4448718, | Sep 13 1983 | Massachusetts Institute of Technology; MASSACHUSETTS INSTITUTE OF TECHNOLOGY, 77 MASSACHUSETTS AVE , CAMBRIDGE, MA 02139 A MA CORP | Method for the preparation of collagen-glycosaminoglycan composite materials |
4451397, | Nov 30 1981 | CENTRE TECHNIQUE CUIR, CHAUSSURE, MAROQUINERIE | Method of preparing collagen products |
4585754, | Jan 09 1984 | FLS ACQUISITION CORP , D B A BIORELEASE TECHNOLOGIES | Stabilization of proteins and peptides by chemical binding with chondroitin |
4703108, | Mar 27 1984 | University of Medicine & Dentistry of New Jersey | Biodegradable matrix and methods for producing same |
4745098, | Feb 24 1984 | The Regents of the University of California; University of California | Compositions and method for improving wound healing |
4751935, | Apr 09 1987 | LEE PHARMACEUTICALS, 1444 SANTA ANITA AVE , SOUTH EL MONTE, CA 91733, A CORP OF CA | Artificial fingernail |
4760131, | Apr 23 1986 | CELTRIX PHARMACEUTICALS, INC | Wound-healing composition |
4808570, | Feb 24 1984 | University of California; REGENTS OF THE UNIVERSITY OF CALIFORNIA THE | Compositions and method for improving wound healing |
4837285, | Mar 27 1984 | MediMatrix | Collagen matrix beads for soft tissue repair |
4880429, | Jul 20 1987 | REGEN CORPORATION, | Prosthetic meniscus |
4947840, | Aug 21 1987 | MASSACHUSETTS INSTITUTE OF TECHNOLOGY, 77 MASSACHUSETTS AVENUE, CAMBRIDGE, MASSACHUSETTS 02139, A CORP OF MA | Biodegradable templates for the regeneration of tissues |
5007934, | Jul 20 1987 | ReGen Corporation | Prosthetic meniscus |
5008116, | Nov 14 1988 | Biomaterials Corporation | Immunostimulatory microsphere |
5028597, | Apr 07 1986 | DAIICHI PHARMACEUTICAL CO , LTD | Antithrombogenic materials |
5071436, | Jul 30 1985 | BASF Beauty Care Solutions France SAS | Substitute bony matrix products promoting osteogenesis |
5108438, | Jul 20 1987 | ReGen Corporation | Prosthetic intervertebral disc |
5116374, | Jul 20 1987 | ReGen Corporation | Prosthetic meniscus |
5116824, | May 02 1985 | Katakura Chikkarin Co., Ltd.; Koken Co., Ltd. | Biomaterial comprising a composite material of a chitosan derivative and collagen derivative |
5158574, | Jul 20 1987 | ReGen Corporation | Prosthetic meniscus |
5169631, | Oct 10 1989 | C S ; LABORATOIRES C S | Topical antimicrobial composition and uses |
5209776, | Jul 27 1990 | TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF YORK, THE | Tissue bonding and sealing composition and method of using the same |
5258043, | Jul 20 1987 | ReGen Corporation | Method for making a prosthetic intervertebral disc |
5263984, | Jul 20 1987 | REGEN BIOLOGICS, INC | Prosthetic ligaments |
5273900, | Apr 28 1987 | The Regents of the University of California | Method and apparatus for preparing composite skin replacement |
5282859, | Apr 24 1990 | ORTEC INTERNATIONAL, INC | Composite living skin equivalents |
5306311, | Jul 20 1987 | ReGen Corporation | Prosthetic articular cartilage |
5605938, | May 31 1991 | TRIAD, LLC | Methods and compositions for inhibition of cell invasion and fibrosis using dextran sulfate |
5624463, | Mar 02 1989 | REGEN BIOLOGICS, INC | Prosthetic articular cartilage |
5629191, | Jan 03 1985 | INTEGRA LIFESCIENCES CORPORATION | Method of making a porous matrix particle |
5645591, | May 29 1990 | MARIEL THERAPEUTICS, INC | Synthetic bone matrix |
5681353, | Jul 20 1987 | REGEN BIOLOGICS, INC | Meniscal augmentation device |
5705177, | May 31 1991 | TRIAD, LLC | Methods and compositions based on inhibition of cell invasion and fibrosis by anionic polymers |
5705178, | Dec 08 1993 | TRIAD, LLC | Methods and compositions based on inhibition of cell invasion and fibrosis by anionic polymers |
5714582, | Mar 17 1995 | BIOSCIENCE CONSULTANTS, L L C | Invertebrate type V telopeptide collagen, methods of making, and use thereof |
5731298, | Jan 03 1992 | Method for the treatment of scars and keloids | |
5735902, | Jul 20 1987 | REGEN BIOLOGICS, INC | Hand implant device |
5735903, | Jul 20 1987 | Ivy Sports Medicine, LLC | Meniscal augmentation device |
5814328, | Jan 13 1997 | Preparation of collagen using papain and a reducing agent | |
5916557, | Nov 12 1993 | TARRANT, LAURENCE J B ; TOKUMO, TOSHIMASA | Methods of repairing connective tissues |
5994325, | May 31 1991 | TRIAD, LLC | Methods and compositions based on inhibition of cell invasion and fibrosis by anionic polymers |
6020326, | May 31 1991 | TRIAD, LLC | Method for inhibition of bone growth by anionic polymers |
6042610, | Jul 20 1987 | REGEN BIOLOGICS, INC | Meniscal augmentation device |
6083930, | May 31 1991 | TRIAD, LLC | Methods and compositions based on inhibition of cell invasion and fibrosis by anionic polymers |
6127348, | Jun 06 1995 | TRIAD, LLC | Methods and compositions based on inhibition of cell invasion and fibrosis by anionic polymers |
6468308, | May 29 1990 | Stryker Corporation | Synthetic bone matrix |
6548077, | Jan 13 1997 | Purifying type I collagen using two papain treatments and reducing and delipidation agents | |
6605117, | May 29 1990 | Stryker Corporation | Synthetic bone matrix |
6916910, | Mar 17 1995 | Bioscience Consultants | Method and process for the production of collagen preparations from invertebrate marine animals and compositions thereof |
6933326, | Jun 19 1998 | LifeCell Corporation | Particulate acellular tissue matrix |
6982298, | Jan 10 2003 | The Cleveland Clinic Foundation; CLEVELAND CLINIC FOUNDATION, THE | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
7309359, | Aug 21 2003 | Warsaw Orthopedic, Inc | Allogenic/xenogenic implants and methods for augmenting or repairing intervertebral discs |
7314860, | Nov 25 1997 | Jenny Ja Antti Wilhurin Rahasto | Heparin-like compounds, their preparation and use to prevent arterial thrombosis associated with vascular injury and interventions |
7358284, | Jun 19 1998 | LifeCell Corporation | Particulate acellular tissue matrix |
7368502, | Jan 08 2004 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
7465766, | Jan 08 2004 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
7504113, | Nov 25 1997 | Jenny Ja Antti Wihurin Rahasto | Heparin-like compounds, their preparation and use to prevent arterial thrombosis associated with vascular injury and interventions |
7713303, | Sep 18 2002 | Warsaw Orthopedic, Inc | Collagen-based materials and methods for augmenting intervertebral discs |
7731981, | Nov 15 2002 | Warsaw Orthopedic, Inc. | Collagen-based materials and methods for treating synovial joints |
7744651, | Sep 18 2002 | Warsaw Orthopedic, Inc | Compositions and methods for treating intervertebral discs with collagen-based materials |
7887593, | Sep 18 2002 | Warsaw Orthopedic, Inc | Method of implanting natural tissue within the vertebral disc nucleus space using a drawstring |
8021350, | Jan 10 2003 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
8070827, | Jul 03 2007 | OCUGEN, INC | Method for use of a double-structured tissue implant for treatment of tissue defects |
8080260, | Feb 13 2008 | The Cleveland Clinic Foundation | Molecular enhancement of extracellular matrix and methods of use |
8118779, | Jun 30 2006 | Warsaw Orthopedic, Inc | Collagen delivery device |
8137688, | Jan 10 2003 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
8138265, | Jan 10 2003 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
8153591, | Aug 26 2004 | PETVIVO HOLDINGS, INC | Protein biomaterials and biocoacervates and methods of making and using thereof |
8207262, | Jan 10 2003 | The Cleveland Clinic Foundation | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
8399619, | Jun 30 2006 | Warsaw Orthopedic, Inc | Injectable collagen material |
8410180, | Apr 30 2008 | The Cleveland Clinic Foundation | Methods to treat urinary incontinence |
8415324, | Nov 25 1997 | Jenny Ja Antti Wihurin Rahasto | Heparin-like compounds, their preparation and use to prevent arterial thrombosis associated with vascular injury and interventions |
8465537, | Jun 17 2003 | PETVIVO HOLDINGS, INC | Encapsulated or coated stent systems |
8623393, | Apr 29 2002 | PETVIVO HOLDINGS, INC | Biomatrix structural containment and fixation systems and methods of use thereof |
8685107, | Jul 03 2007 | OCUGEN, INC | Double-structured tissue implant and a method for preparation and use thereof |
8871267, | Sep 25 1998 | PETVIVO HOLDINGS, INC | Protein matrix materials, devices and methods of making and using thereof |
8921109, | Sep 19 2005 | OCUGEN, INC | Cell-support matrix having narrowly defined uniformly vertically and non-randomly organized porosity and pore density and a method for preparation thereof |
9011543, | Aug 21 2003 | Warsaw Orthopedic, Inc. | Methods for augmenting or repairing intervertebral discs with allogenic/xenogenic implants |
9107937, | Aug 26 2003 | PETVIVO HOLDINGS, INC | Wound treatments with crosslinked protein amorphous biomaterials |
9149562, | Jul 03 2007 | OCUGEN, INC | Method for use of a double-structured tissue implant for treatment of tissue defects |
9271821, | Jan 24 2012 | LifeCell Corporation | Elongated tissue matrices |
9393347, | Jul 03 2007 | OCUGEN, INC | Double-structured tissue implant and a method for preparation and use thereof |
9421304, | Jul 03 2007 | OCUGEN, INC | Method for improvement of differentiation of mesenchymal stem cells using a double-structured tissue implant |
9532863, | Dec 20 2011 | LifeCell Corporation | Sheet tissue products |
9549805, | Dec 20 2011 | LifeCell Corporation | Flowable tissue products |
9592125, | Dec 22 2006 | LABORATOIRE MEDIDOM S A | In situ system for intra-articular chondral and osseous tissue repair |
9687590, | Jul 03 2007 | OCUGEN, INC | Double-structured tissue implant and a method for preparation and use thereof |
9701940, | Sep 19 2005 | OCUGEN, INC | Cell-support matrix having narrowly defined uniformly vertically and non-randomly organized porosity and pore density and a method for preparation thereof |
9913705, | Dec 20 2011 | LifeCell Corporation | Flowable tissue products |
9993326, | Jul 03 2007 | OCUGEN, INC | Method for use of a double-structured tissue implant for treatment of tissue defects |
9999637, | Apr 24 2012 | LifeCell Corporation | Functionalized tissue matrices |
9999705, | Aug 26 2003 | Gel-Del Technologies, Inc. | Protein biomaterials and biocoacervates and methods of making and using thereof |
RE35399, | Apr 24 1990 | ORCEL LLC | Composite living skin equivalents |
Patent | Priority | Assignee | Title |
3527225, | |||
4060081, | Jul 15 1975 | Massachusetts Institute of Technology | Multilayer membrane useful as synthetic skin |
4233360, | Nov 22 1975 | COHESION TECHNOLOGIES, INC | Non-antigenic collagen and articles of manufacture |
4280954, | Jul 15 1975 | Massachusetts Institute of Technology | Crosslinked collagen-mucopolysaccharide composite materials |
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